Expression Of Nitrate Reductase Research Articles

StCDF1: A 'jack of all trades' clock output with a central role in regulating potato nitrate reduction activity.

Transcription factors of the CYCLING DOF FACTOR (CDF) family activate in potato the SP6A FT tuberization signal in leaves. In modern cultivars, truncated StCDF1.2 alleles override strict SD control by stabilizing the StCDF1 protein, which leads to StCOL1 suppression and impaired activation of the antagonic SP5G paralog. By using DAP-seq and RNA-seq studies, we here show that StCDF1 not only acts as an upstream regulator of the day length pathway but also directly regulates several N assimilation and transport genes. StCDF1 directly represses expression of NITRATE REDUCTASE (NR/NIA), which catalyses the first reduction step in nitrate assimilation, and is encoded by a single potato locus. StCDF1 knock-down lines performed better in N-limiting conditions, and this phenotype correlated with derepressed StNR expression. Also, deletion of the StNR DAP-seq region abolished repression by StCDF1, while it did not affect NLP7-dependent activation of the StNR promoter. We identified multiple nucleotide polymorphisms in the DAP-seq region in potato cultivars with early StCDF1 alleles, suggesting that this genetic variation was selected as compensatory mechanism to the negative impact of StCDF1 stabilization. Thereby, directed modification of the StCDF1-recognition elements emerges as a promising strategy to enhance limiting StNR activity in potato.

Effect of copper on nitrogen uptake, transportation, assimilation processes, and related gene expression in Chinese cabbage [Brassica campestris L. ssp. Chinensis (L.)] under various nitrate-to-ammonium ratios.

Improving vegetable yield and optimizing its quality through nutrient management have long been central to plant nutrition and horticultural science. Copper (Cu) is recognized as an essential trace element that promotes plant growth and development. However, the mechanisms by which Cu influences nitrogen (N) metabolism remain largely unknown, with limited studies exploring the interaction between Cu and varying nitrate-to-ammonium (nitrate/ammonium) ratios. In this study, Chinese cabbage was exposed to two Cu concentrations (0 and 0.02 mg L-1) in combination with three nitrate/ammonium ratios (10/90, 50/50, and 90/10) under hydroponic conditions. The results showed that Cu application increased plant biomass, nitrate reductase (NR) and glutamine synthetase (GS) enzyme activities, the expression of NR (NIA) and GS2 (Gln2) genes, and N content in both shoots and roots. Additionally, Cu treatment decreased nitrate and free amino acid contents, as well as the expression of nitrate transporters NRT1.1 and NRT2.1 in roots while increasing these four parameters in shoots. Additionally, these effects were significantly modulated by the nitrate/ammonium ratios. In conclusion, Cu may facilitate nitrate transportation, enhance nitrate reduction, promote ammonium assimilation, and influence the transformation of organic N compounds, highlighting its potential role in improving N metabolism in Chinese cabbage.

Whole-Genome Sequencing of Three Lactiplantibacillus plantarum Strains Reveals Potential Metabolites for Boosting Host Immunity Safely.

In response to the growing demand for immune-related products, this study evaluated the safety and immune-modulating potential of three newly discovered Lactiplantibacillus plantarum strains (GKM3, GKK1, and GKD7) through toxicity tests and whole-genome sequencing. Safety evaluations, including the analysis of antimicrobial resistance genes, virulence factors, plasmids, and prophages, classified these strains as safe for human consumption. Acute oral toxicity tests further supported their safety. To evaluate their immune-modulating potential, dendritic cells were exposed to these strains, and the secretion of key cytokines (IFN-β and IL-12) was measured. Among the strains, GKK1 exhibited the highest enhancement of IFN-β and IL-12 production, suggesting its potential as an immune-stimulating probiotic. Bioinformatics analysis revealed potential metabolic pathways and secondary metabolites, including predicted bacteriocins, associated with immune modulation. The presence of a nitrate reductase region in the GKK1 strain indicated its ability to produce nitric oxide, a critical molecule involved in immune regulation and host defense. The presence of glucorhamnan-related gene clusters in GKK1 also suggested immune-enhancing effects. Nitrate reductase expression was confirmed using qPCR, with the highest levels detected in GKK1. Moreover, this study is the first to show an anti-inflammatory effect of plantaricin A, linked to its presence in strain GKM3 and its potential therapeutic applications due to sequence similarity to known anti-inflammatory peptides. Overall, these three L. plantarum strains demonstrated a safe profile and GKK1 showed potential as an immunity-enhancing probiotic. However, additional investigation is required to confirm the involvement of specific metabolic pathways, secondary metabolites, and bacteriocins in immune responses.

Microplastic exposure inhibits nitrate uptake and assimilation in wheat plants

Microplastic (MP) contamination in soil severely impairs plant growth. However, mechanisms underlying the effects of MPs on plant nutrient uptake remain largely unknown. In this study, we revealed that NO3− content was significantly decreased in shoots and roots of wheat plants exposed to high concentrations (50–100 mg L−1) of MPs (1 μm and 0.1 μm; type: polystyrene) in the hydroponic solution. Isotope labeling experiments demonstrated that MP exposure led to a significant inhibition of NO3− uptake in wheat roots. Further analysis indicated that the presence of MPs markedly inhibited root growth and caused oxidative damage to the roots. Additionally, superoxide dismutase and peroxidase activities in wheat roots decreased under all MP treatments, whereas catalase and ascorbate peroxidase activities significantly increased under the 100 mg L−1 MP treatment. The transcription levels of most nitrate transporters (NRTs) in roots were significantly downregulated by MP exposure. Furthermore, exposure to MPs distinctly suppressed the activity of nitrate reductase (NR) and nitrite reductase (NiR), as well as the expression levels of their coding genes in wheat shoots. These findings indicate that a decline in root uptake area and root vitality, as well as in the expression of NRTs, NR, and NiR genes caused by MP exposure may have adverse effects on NO3− uptake and assimilation, consequently impairing normal growth of plants.

Proline interacts with Ca2+-dependent signaling to enhance chromium tolerance in rice by manipulating nitrate reductase and sucrose phosphate synthase

The entrance of chromium (Cr) into the agricultural system would exert a negative influence on the carbon/nitrogen metabolism (CNM) of plants. In this study, we investigated the role of exogenous proline-mediated Ca2+-dependent signaling in the regulation of CNM in rice subjected to Cr(VI) stress, with emphasis on the involvement of nitrate reductase (NR) and sucrose phosphate synthase (SPS). Results demonstrated that proline effectively mitigated the growth inhibition of rice imposed by Cr(VI) stress, which is achieved by a reduction in cytoplasmic Ca and Cr content and the activation of the downstream Ca2+-dependent signaling pathway. Additionally, proline displayed a positive effect in modulating the expression and activities of NR and SPS under Cr(VI) stress, which are attributed to the cross-regulation between calcium-dependent protein kinases (CDPKs) and 14-3-3 proteins (14-3-3s). Consequently, nitrogen use efficiency and sucrose content in rice under Cr(VI) + proline treatments were higher than Cr(VI) treatments. Gene expression variation factors underscored that the regulation of proline on NR is crucial to the Ca2+-dependent signaling pathway, initiated by the interaction between CDPKs and 14-3-3s in rice plants during Cr(VI) stress. These results reveal that proline interacts with Ca2+-dependent signaling pathways to enhance Cr tolerance in rice by regulating NR and SPS.

Lack of methoxy-mycolates characterizes the geographically restricted lineage 7 of Mycobacterium tuberculosis complex.

Lineage 7 (L7) emerged in the phylogeny of the Mycobacterium tuberculosis complex (MTBC) subsequent to the branching of 'ancient' lineage 1 and prior to the Eurasian dispersal of 'modern' lineages 2, 3 and 4. In contrast to the major MTBC lineages, the current epidemiology suggests that prevalence of L7 is highly confined to the Ethiopian population, or when identified outside of Ethiopia, it has mainly been in patients of Ethiopian origin. To search for microbiological factors that may contribute to its restricted distribution, we compared the genome of L7 to the genomes of globally dispersed MTBC lineages. The frequency of predicted functional mutations in L7 was similar to that documented in other lineages. These include mutations characteristic of modern lineages - such as constitutive expression of nitrate reductase - as well as mutations in the VirS locus that are commonly found in ancient lineages. We also identified and characterized multiple lineage-specific mutations in L7 in biosynthesis pathways of cell wall lipids, including confirmed deficiency of methoxy-mycolic acids due to a stop-gain mutation in the mmaA3 gene that encodes a methoxy-mycolic acid synthase. We show that the abolished biosynthesis of methoxy-mycolates of L7 alters the cell structure and colony morphology on selected growth media and impacts biofilm formation. The loss of these mycolic acid moieties may change the host-pathogen dynamic for L7 isolates, explaining the limited geographical distribution of L7 and contributing to further understanding the spread of MTBC lineages across the globe.

Effects of nitrate and ammonium on assimilation of nitric oxide by Heterosigma akashiwo

The harmful alga Heterosigma akashiwo possesses a hybrid nitrate reductase (NR) enzyme, NR2-2/2HbN, which has the potential to convert NO to nitrate for assimilation into biomass. In previous research, NR transcription in H. akashiwo was induced by nitrate while NR activity was inhibited by ammonium. Here, the capacity of H. akashiwo to use NO in the presence of nitrate and/or ammonium was investigated to understand the regulation of NO assimilation. Continuous cultures of H. akashiwo were acclimated to growth on nitrate, ammonium, or a mixture of both. Aliquots from these cultures were spiked with 15N-labeled NO. The expression of genes involved in nitrogen assimilation was evaluated, as well as nitrate reductase activity and assimilation of 15N-labeled nitrogen into algal biomass. Results showed that NO induced expression and activity of NR, and upregulated expression of GOGAT regardless of the presence of other inorganic nitrogen sources, while GS expression decreased over time. Furthermore, 15NO uptake and assimilation was significantly higher in cultures acclimated for growth on ammonium compared to cultures acclimated for growth on nitrate alone. Assimilation of NO may provide H. akashiwo with a competitive advantage in N-poor environments or areas with elevated NO.

Key role of reactive oxygen species-scavenging system in nitric oxide and hydrogen sulfide crosstalk-evoked thermotolerance in maize seedlings.

Nitric oxide (NO) and hydrogen sulfide (H2S) are novel signaling molecules, which participate in plant growth, development, and response to stress. In this study root-irrigation with 0.15 mM sodium nitroprusside (SNP, NO donor) up-regulated gene expression of L-CYSTEINE DESULFHYDRASE1 (LCD1), activities of L-cysteine desulfhydrase (LCD) and D-cysteine desulfhydrase (DCD), as well as an endogenous H2S level, compared to control seedlings. The SNP-up-regulated effects were enhanced by 0.5 mM sodium hydrosulfide (NaHS, H2S donor), but weakened by NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and H2S scavenger hypotaurine (HT) alone. NaHS had no significant effect on gene expression and activity of nitrate reductase (NR, a NO candidate producing enzyme). These data indicate that NO could trigger the LCD/H2S signaling pathway in maize seedlings. To further investigate the effect of NO and H2S crosstalk on thermotolerance in maize seedlings, thermotolerance parameters and reactive oxygen species (ROS)-scavenging system were estimated. The results show that SNP increased survival rate and tissue viability, decreased malondialdehyde (MDA) accumulation, and electrolyte leakage in maize seedlings under heat stress (HS), implying NO could improve thermotolerance in maize seedlings. The NO-improved thermotolerance was impaired by H2S inhibitor DL-propargylglycine (PAG) and scavenger HT alone. Similarly, SNP up-regulated the gene expression of DEHYDROASCORBATE REDUCTASE (DHAR) and GLUTATHIONE REDUCTASE1 (GR1); activities of ascorbate peroxidase, glutathione reductase, and catalase; as well as levels of ascorbic acid, glutathione, flavonoids, carotenoids, and total phenols. SNP also reduced hydrogen peroxide and superoxide radical accumulation in maize seedlings under HS compared to the control. The effects of SNP on ROS and their scavenger system were weakened by PAG and HT alone. These data hint that NO could evoke thermotolerance in maize seedlings by triggering the LCD/H2S signaling pathway, and the ROS-scavenging system played a key role in the NO and H2S crosstalk-evoked thermotolerance.

Moderately Reducing Nitrogen Application Ameliorates Salt-Induced Growth and Physiological Damage on Forage Bermudagrass.

Nitrogen (N) application is one of the most effective methods to alleviate salt-induced damage on plants. Forage bermudagrass has higher utilization potential on saline soil, but whether its N requirement changed under high salt stress has not been studied. Through examining plant growth-related traits, salt-stress-responsive physiological traits, photosynthesis, N metabolism, and forage quality supplied with different N concentrations under high salt stress (200 mM NaCl), we noticed that the optimum N requirement of forage bermudagrass reduced. When supplied with 10 mM N under higher salt stress, plants had a similar biomass, turf color, and chlorophyll content with plants supplied with 15 mM N, accompanied by a lower firing rate and Na+ content of leaves. The N content, crude protein, crude fat content, the expression of AMTs (ammonium transporters), NR (nitrate reductase), GS (glutamine synthetase), and GOGAT (glutamate synthetase), the chlorophyll fluorescence curve, and parameters of leaves (e.g., PIABS; PICS; ABS/RC; TRo/RC; ETo/RC) all peaked under 10 mM N under high salt stress instead of 15 mM N. Through exploring the proper N application under higher salt stress and its alleviation mechanisms, our results indicated that moderate reduction in N application under high salt level had a maximum promotion effect on the salt tolerance of forage bermudagrass without growth or forage quality inhibition. These response mechanisms obtained can provide a useful reference for N application in moderation rather than in excess on forage bermudagrass, especially in higher salinity areas.

New insights into the role of chrysanthemum calcineurin B\u2013like interacting protein kinase CmCIPK23 in nitrate signaling in Arabidopsis roots

Nitrate is an important source of nitrogen and also acts as a signaling molecule to trigger numerous physiological, growth, and developmental processes throughout the life of the plant. Many nitrate transporters, transcription factors, and protein kinases participate in the regulation of nitrate signaling. Here, we identified a gene encoding the chrysanthemum calcineurin B-like interacting protein kinase CmCIPK23, which participates in nitrate signaling pathways. In Arabidopsis, overexpression of CmCIPK23 significantly decreased lateral root number and length and primary root length compared to the WT when grown on modified Murashige and Skoog medium with KNO3 as the sole nitrogen source (modified MS). The expression of nitrate-responsive genes differed significantly between CmCIPK23-overexpressing Arabidopsis (CmCIPK23-OE) and the WT after nitrate treatment. Nitrate content was significantly lower in CmCIPK23-OE roots, which may have resulted from reduced nitrate uptake at high external nitrate concentrations (≥ 1 mM). Nitrate reductase activity and the expression of nitrate reductase and glutamine synthase genes were lower in CmCIPK23-OE roots. We also found that CmCIPK23 interacted with the transcription factor CmTGA1, whose Arabidopsis homolog regulates the nitrate response. We inferred that CmCIPK23 overexpression influences root development on modified MS medium, as well as root nitrate uptake and assimilation at high external nitrate supply. These findings offer new perspectives on the mechanisms by which the chrysanthemum CBL interacting protein kinase CmCIPK23 influences nitrate signaling.

Autophagic pathway contributes to low-nitrogen tolerance by optimizing nitrogen uptake and utilization in tomato.

Autophagy is a primary process involved in the degradation and reuse of redundant or damaged cytoplasmic components in eukaryotes. Autophagy has been demonstrated to facilitate nutrient recycling and remobilization by delivering intracellular materials to the vacuole for degradation in plants under nutrient starvation. However, the role of autophagy in nitrogen (N) uptake and utilization remains unknown. Here, we report that the ATG6-dependent autophagic pathway regulates N utilization in tomato (Solanum lycopersicum) under low-nitrogen (LN) conditions. Autophagy-disrupted mutants exhibited weakened biomass production and N accumulation compared with wild-type (WT), while ATG6 overexpression promoted autophagy and biomass production under LN stress. The N content in atg6 mutants decreased while that in ATG6-overexpressing lines increased due to the control of N transporter gene expression in roots under LN conditions. Furthermore, ATG6-dependent autophagy enhanced N assimilation efficiency and protein production in leaves. Nitrate reductase and nitrite reductase activities and expression were compromised in atg6 mutants but were enhanced in ATG6-overexpressing plants under LN stress. Moreover, ATG6-dependent autophagy increased plant carbon fixation and photosynthetic capacity. The quantum yield of photosystem II, photosynthetic N use efficiency and photosynthetic protein accumulation were compromised in atg6 mutants but were restored in ATG6-overexpressing plants. A WT scion grafted onto atg6 mutant rootstock and an atg6 scion grafted onto WT rootstock both exhibited inhibited LN-induced autophagy and N uptake and utilization. Thus, ATG6-dependent autophagy regulates not only N uptake and utilization as well as carbon assimilation but also nutrient recycling and remobilization in tomato plants experiencing LN stress.

Nitrogen nutrition modifies the susceptibility of Arabidopsis thaliana to the necrotrophic fungus, Alternaria brassicicola.

The impact of the form of nitrogen (N) source (nitrate versus ammonium) on the susceptibility to Alternaria brassicicola, a necrotrophic fungus, has been examined in Arabidopsis thaliana at the rosette stage. Nitrate nutrition was found to increase fungal lesions considerably. There was a similar induction of defence gene expression following infection under both N nutritions, except for the phytoalexin deficient 3 gene, which was overexpressed with nitrate. Nitrate also led to a greater nitric oxide production occurring in planta during the saprophytic growth and lower nitrate reductase (NIA1) expression 7 days after inoculation. This suggests that nitrate reductase-dependent nitric oxide production had a dual role, whereby, despite its known role in the generic response to pathogens, it affected plant metabolism, and this facilitated fungal infection. In ammonium-grown plants, infection with A. brassicicola induced a stronger gene expression of ammonium transporters and significantly reduced the initially high ammonium content in the leaves. There was a significant interaction between N source and inoculation (presence versus absence of the fungus) on the total amino acid content, while N nutrition reconfigured the spectrum of major amino acids. Typically, a higher content of total amino acid, mainly due to a stronger increase in asparagine and glutamine, is observed under ammonium nutrition while, in nitrate-fed plants, glutamate was the only amino acid which content increased significantly after fungal inoculation. N nutrition thus appears to control fungal infection via a complex set of signalling and nutritional events, shedding light on how nitrate availability can modulate disease susceptibility.

Nitrogen fertilization stimulates nitrogen assimilation and modifies nitrogen partitioning in the spring shoot leaves of citrus (Citrus reticulata Blanco) trees

The spring shoot leaves are important sites of nitrogen (N) metabolism in citrus trees. Understanding the physiological and metabolic response of the spring shoot leaves under varying N fertilization is fundamental to the fertilization management in citrus orchards. Thus, the processes affecting N composition, the activities of N metabolism related enzymes, and the expression of relevant genes were explored in spring shoot leaves under four N levels (0, 207, 275, 413 g N tree−1 y−1, as N0, N207, N275, N413). The results showed that, compared with N0, N275 significantly increased total N by 24.81%, which was mainly attributed to enhancement of structural N by 30.92%, free amino acid N by 40.91% and nitrate N by 41.33%. The relative expression of nitrate reductase (NR) and glutamate dehydrogenase (GDH) under N275 increased by 19.32% and 73.48%, respectively, compared with that under N0 treatment. Compared with N0 treatment, the NR transcription level under N275 treatment increased by 381%. The relative transcription levels of NADP-GDH and GDH1 also increased with increasing N fertilization. However, compared with that under N275, the relative transcription of GDH2 under N413 treatment was inhibited. Therefore, the transcript abundance of NR, NADP-GDH,GDH1 and GDH2 affected the activities of NR and GDH and thereby contributed to the regulation of N composition in the leaves. In addition, the activities of glutamine synthetase and nitrite reductase were largely unaffected or even declined in the N207, N275 and N413 treatments compared with the N0. This study elucidated the mechanism of primary N metabolism and partitioning in citrus leaves and provided a theoretical basis for N management in citrus orchards.

Increased nitrate concentration differentially affects cell growth and expression of nitrate transporter and other nitrogen-related genes in the harmful dinoflagellate Prorocentrum minimum

The molecular mechanisms through which dinoflagellates adapt to nitrate fluctuations in aquatic environments remain poorly understood. Here, we sequenced the full-length cDNA of a nitrate transporter (NRT) gene from the harmful marine dinoflagellate Prorocentrum minimum Schiller. The cDNA length was 2431 bp. It encoded a 529-amino acid protein, which was phylogenetically clustered with proteins from other dinoflagellates. Nitrate supply promoted cell growth up to a certain concentration (∼1.76 mM) but inhibited it at higher concentrations. Interestingly, at the inhibitory concentrations, nitrite levels in the medium were considerably increased. Nitrate concentration affected the expression of PmNRT, nitrite transporter (PmNiRT), nitrate reductase (PmNR), and nitrite reductase (PmNiR). Specifically, PmNRT was upregulated after 24 h, with ∼6-fold change compared with the control level, in both nitrate-depleted and nitrate-repleted cultures. In addition, PmNR transcript levels increased to the maximum of 4-fold at 48 h but decreased thereafter. In contrast, PmNiR levels remained unchanged in both nitrate-repleted and nitrate-depleted cultures. Therefore, P. minimum likely copes with nitrate fluctuations in its environment by regulating a set of genes responsible for nitrate uptake.

New insights into N-utilization efficiency in tomato (Solanum lycopersicum L.) under N limiting condition

Understanding Nitrogen Use Efficiency (NUE) physiological and molecular mechanisms in high N demanding crops has become decisive for improving NUE in sustainable cropping systems. How the Nitrogen Utilization Efficiency (NUtE) component contributes to the NUE enhancement under nitrate limiting conditions in tomato remains to be elucidated. This study deals with the changes in several important nitrate metabolism related gene expressions (nitrate assimilation, transport, remobilization and storage/sequestration) engendered by short and long-term limiting nitrate exposure in two selected NUE-contrasting genotypes, Regina Ostuni (RO) and UC82, efficient and inefficient, respectively. At short-term, nitrate limiting supply triggered higher SlCLCa and SlNRT1.7 expressions in RO root and shoot, respectively, suggesting a higher nitrate storage and remobilization compared to UC82, explaining how RO withstood the nitrate deficiency better than UC82. At long-term, nitrate reductase (SlNR) and nitrite reductase (SlNIR) expression were not significantly different between nitrate treatments in RO, while significantly down-regulated under nitrate limiting treatment in UC82. In addition, SlCLCa and SlNRT1.8 transcript levels were significantly lower in RO, while those of SlNRT1.5 and SlNR appeared significantly higher. This suggested that the efficient genotype stored less nitrate compared to UC82, which was allocated and assimilated to the shoot. More interestingly, the expression of SlNRT2.7 was significantly higher in RO shoot compared to UC82 and strongly correlated to RO higher growth as well as to NUE and NUtE component. Our findings underlined the differential regulation of N-metabolism genes that may confer to NUtE component a pivotal role in NUE enhancement in tomato.

Barley transcription factor HvNLP2 mediates nitrate signaling and affects nitrogen use efficiency.

Plants have evolved complex mechanisms to adapt to the changing nitrogen levels in the environment. In Arabidopsis, more than a dozen nitrate signaling regulatory genes have been characterized, including the NODULE INCEPTION-LIKE PROTEIN (AtNLP) genes, which play essential roles in nitrate signaling. However, whether NLP genes in the Triticeae crops are involved in nitrate regulation and nitrogen use efficiency (NUE) remains unknown. Here, we isolated a barley (Hordeum vulgare L.) mutant, hvnlp2-1, from a TILLING (Targeting Local Lesions IN Genomes) population and constructed two RNAi lines, hvnlp2-2 and hvnlp2-3, to study the function of HvNLP2. The expression of the nitrate-responsive genes was substantially inhibited after nitrate treatment in the hvnlp2 mutants, indicating that HvNLP2 controls nitrate signaling. Nitrate content was significantly higher in the hvnlp2 mutants, which may result from the decreased assimilation of nitrogen caused by reduced nitrate reductase activity and expression of nitrate assimilatory genes. HvNLP2 is localized to the nucleus in the presence of nitrate. Further investigation showed that HvNLP2 binds to and activates the nitrate-responsive cis-elements. Moreover, hvnlp2 exhibited reduced biomass, seed yield, and NUE. Therefore, HvNLP2 controls nitrate signaling and plays an important role in NUE.

Expression dynamics of genes encoding nitrate and ammonium assimilation enzymes in rice genotypes exposed to reproductive stage salinity stress

Understanding the reproductive stage salinity stress tolerance is a key target for breeding stress tolerant rice genotypes. Nitrate and ammonium are equally important nitrogen forms utilized by rice. We evaluated nitrate and ammonium assimilation during reproductive stage in control and salinity (10dSm−1 using NaCl) stressed rice plants. Osmotic stress tolerant rice genotype Shabhagidhan (SD) and high yielding yet osmotic and salinity stress sensitive genotype Pusa sugandh-5 (PS5) were evaluated. Salinity stress was given to plants during panicle emergence and flag leaves was collected after 1d, 3d 5d, 7d, 9d,12d and 15d after anthesis. Reproductive stage salinity stress resulted in decrease of membrane stability, relative water content and osmotic potential of rice plants. Reproductive stage salinity stress decreased the expression of nitrate reductase (OsNIA), nitrite reductase (OsNiR), Glutamine synthetase (OsGLN1.1, OsGLN1.2, OsGLN2) and glutamate synthase/GOGAT (OsFd-GOGAT, OsNADH-GOGAT) in flag leaves. In response to stress, SD showed better stress tolerance than PS5 in terms of higher yield stability. Variety SD showed higher leaf nitrate and ammonium content and maintained comparatively higher nitrate and ammonia metabolism enzyme activity than PS5. Salinity stress upregulated the activity of glutamate dehydrogenase enzyme and indirectly contributed to the higher proline content and maintenance of favourable osmotic potential in SD. Expression of GS2 which has role in photo respiratory ammonia assimilation was upregulated by salinity stress in PS5 in comparison to SD. Rice genotype showing better induction of nitrogen assimilatory genes will be more tolerant to reproductive stage salinity stress.

Denitrifying bacteria respond to and shape microscale gradients within particulate matrices

Heterotrophic denitrification enables facultative anaerobes to continue growing even when limited by oxygen (O2) availability. Particles in particular provide physical matrices characterized by reduced O2 permeability even in well-oxygenated bulk conditions, creating microenvironments where microbial denitrifiers may proliferate. Whereas numerical particle models generally describe denitrification as a function of radius, here we provide evidence for heterogeneity of intraparticle denitrification activity due to local interactions within and among microcolonies. Pseudomonas aeruginosa cells and microcolonies act to metabolically shade each other, fostering anaerobic processes just microns from O2-saturated bulk water. Even within well-oxygenated fluid, suboxia and denitrification reproducibly developed and migrated along sharp 10 to 100 µm gradients, driven by the balance of oxidant diffusion and local respiration. Moreover, metabolic differentiation among densely packed cells is dictated by the diffusional supply of O2, leading to distinct bimodality in the distribution of nitrate and nitrite reductase expression. The initial seeding density controls the speed at which anoxia develops, and even particles seeded with few bacteria remain capable of becoming anoxic. Our empirical results capture the dynamics of denitrifier gene expression in direct association with O2 concentrations over microscale physical matrices, providing observations of the co-occurrence and spatial arrangement of aerobic and anaerobic processes.

Synergetic effects of biochars and denitrifier on nitrate removal

Nitrate is one of the most common water contaminants and has caused severe environmental problems. This work aimed to investigate the effects of integration of denitrifier with biochars on nitrate removal and understand the underlying mechanisms. The results showed that physiochemical properties of biochars varied according to different feedstocks, which influenced bacteria attachment and nitrate removal through adsorption. However, bacteria could colonize on biochars no matter biochars surface were favorable for bacteria attachment or not. Immobilization of denitrifier on biochars significantly improved nitrate removal efficiencies and reduced lag time. Underlying mechanisms investigation showed that the integration of denitrifier with biochars had synergetic effects on promoting nitrate removal, which improved not only the expression and activity of nitrate reductase, but also the electron transport system activity.

Physiologic and Transcriptomic Insights into the High Alkali Response of Dunaliella salina

The halophilic unicellular green alga Dunaliella salina is characterized as the distinct tolerance to high salt in the field. Normally the organism is concurrently subjected with salt and alkali stress, but the feature of D. salina in response to high alkali stress was not investigated before. In this study, we used high pH stress to simulate high alkali stress and found D. salina also showed tolerance to high alkali stress, even grown under pH at 11.0, with high cell density and chlorophyll biosynthesis. High-throughput transcriptome analysis revealed the dramatically different expression of genes associated with nitrogen metabolism, lipid and β-carotene biosynthesis after high alkali treatment. Furthermore, physiologic analysis showed that high alkali treatment induced the expression and activity of nitrate reductase (NR), which generated nitric oxide (NO) as the messenger, and ultimately induced the accumulation of lipid and β-carotene biosynthesis. Suppressing NR activity or NO generation also reduced high-alkali-induced lipid and β-carotene biosynthesis, as well as attenuated the tolerance of D. salina to high alkali stress. Thus, our physiologic and transcriptomic data reveal the novel function of NR-dependent NO signal during the adaptation of D. salina to high alkali stress, through inducing the biosynthesis of lipid and β-carotene biosynthesis.